LOCUS       sequence-210782-        2844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP cassette under the control of the ibpAB sigma32-responsive 
            promoter. Used to test burden-responsive behaviour of the wt genomic
            promoter. The plasmid has got ColE1 origin of replication and AmpR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2844)
  AUTHORS   Ceroni F, Boo A, Furini S, Gorochowski TE, Borkowski O, Ladak YN, 
            Awan AR, Gilbert C, Stan GB, Ellis T
  TITLE     Burden-driven feedback control of gene expression.
  JOURNAL   Nat Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635.
  PUBMED    29578536
REFERENCE   2  (bases 1 to 2844)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635."
COMMENT     Sequence Label:  ibpAB-GFP
FEATURES             Location/Qualifiers
     source          1..2844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             96..107
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             114..827
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      839..910
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      926..953
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1077..1868,1869..1937))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1700..1719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1938..2042)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2145..2733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2634..2653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctaggcct gcaaaaaaaa gtccgctgat aaggcttgaa aagttcattt ccagacccat
       61 ttttacatct taattaagga tcttagctac tagagaaaga ggagaaatac tagatgcgta
      121 aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca
      181 acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga
      241 cgctgaagtt catctgtact actggtaaac tgccggttcc ttggccgact ctggtaacga
      301 cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact
      361 tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg
      421 acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca
      481 ttgagctgaa aggcattgac tttaaagagg acggcaatat cctgggccat aagctggaat
      541 acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag
      601 cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc
      661 agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca
      721 cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt
      781 tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatga ccaggcatca
      841 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
      901 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttataa
      961 gtcggtctca tacagcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat
     1021 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1081 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1141 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1201 gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag
     1261 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1321 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1381 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1441 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1501 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1561 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1621 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1681 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1741 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1801 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1861 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1921 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1981 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2041 cgcacatttc cccgaaaagt gccacctgtc atgaccaaaa tcccttaacg tgagttttcg
     2101 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2221 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2281 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2341 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2401 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2461 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2521 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2581 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2641 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2701 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2761 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2821 gtggataacc gtgcggccgc ggcc
//
